Article
Refine
Year of publication
Institute
- Fachbereich Chemie und Biotechnologie (472) (remove)
Language
- English (472) (remove)
Document Type
- Article (472) (remove)
Keywords
- Heparin (3)
- Bacillaceae (2)
- Biotechnological application (2)
- Butanol (2)
- Chemometrics (2)
- IR spectroscopy (2)
- NMR spectroscopy (2)
- Principal component analysis (2)
- Standardization (2)
- Subtilases (2)
- Subtilisin (2)
- bubble column (2)
- methanation (2)
- plug flow reactor (2)
- qNMR (2)
- (Poly)saccharides (1)
- ABE (1)
- Acid crash (1)
- Algal Turf Scrubber (1)
- Algal–bacterial bioflm (1)
- Alginate beads (1)
- Alkalihalobacillus okhensis (1)
- Aloe vera (1)
- Authenticity (1)
- Biofuel (1)
- Biogas (1)
- Biological hydrogen (1)
- Biomass (1)
- Biorefinery (1)
- Bragg peak (1)
- Broad pH spectrum (1)
- C. acetobutylicum (1)
- CRISPR/Cas9 (1)
- Carrier solvents (1)
- Chondroitin sulfate (1)
- Circular bioeconomy (1)
- Clostridium acetobutylicum (1)
- Crude heparin (1)
- Cyclotron production (1)
- Dark fermentation (1)
- Dehydrogenase (1)
- Detergent protease (1)
- Deuterated solvents (1)
- Deuterium NMR (1)
- Diaphorase (1)
- Dietary supplements (1)
- Electronic cigarettes (1)
- Enzymatic biosensor (1)
- Extracellular enzymes (1)
- Free-base nicotine (1)
- Ga-68 (1)
- Glucosamine (1)
- Halotolerant protease (1)
- High-field NMR (1)
- Hydrolysis (1)
- Hypersecretion (1)
- IR (1)
- Inorganic ions (1)
- Ions (1)
- Lignocellulose (1)
- Linear discriminant analysis (1)
- Low field NMR (1)
- Manufacturer (1)
- Marker-free mutagenesis (1)
- Medical radionuclide production (1)
- Metabolic shift (1)
- Metal contaminants (1)
- Methane (1)
- Microfluidic solvent extraction (1)
- Molecular modelling (1)
- Molecular weight determination (1)
- NMR (1)
- Organic acids (1)
- Organic waste (1)
- P2G (1)
- PLS-regression (1)
- Polysaccharides (1)
- Pre-culture (1)
- Pretreatment (1)
- Quality control (1)
- Quantum chemistry (1)
- Simultaneous determination (1)
- Soft independent modeling of class analogy (1)
- Spectroscopy (1)
- Stenotrophomonas maltophilia (1)
- Streptomyces griseus (1)
- Streptomyces lividans (1)
- USP (1)
- Uracil-phosphoribosyltransferase (1)
- Weak organic acids (1)
- acetoin (1)
- acetoin reductase (1)
- actuator-sensor system (1)
- alcoholic beverages (1)
- aspergillus (1)
- bacterial cellulose (1)
- bi-enzyme biosensor (1)
- bio-methane (1)
- bioavailability (1)
- biodegradable polymers (1)
- biological dosimeter (1)
- biomethane (1)
- biosensors (1)
- borehole disposal (1)
- capacitive field-effect sensor (1)
- capacitive field-effect sensors (1)
- coculture (1)
- deficit irrigation (1)
- detergent protease (1)
- disposal facility (1)
- drug metabolising enzymes (1)
- drug–drug interactions (1)
- elastomers (1)
- enzyme kinetics (1)
- enzyme-logic gate (1)
- exopolysaccharides (1)
- filamentous fungi (1)
- genome engineering (1)
- geological disposal (1)
- glycine (1)
- halotolerant protease (1)
- high-alkaline subtilisin (1)
- human metabolites (1)
- hydrogel (1)
- hydrogels (1)
- light-addressable electrode (1)
- light-addressable potentiometric sensor (1)
- mechanical properties (1)
- microfluidics (1)
- micronutrients (1)
- neutrons (1)
- nuclear waste (1)
- onion (1)
- optical fibers (1)
- oxidative stable protease (1)
- penicillinase (1)
- polyaspartic acid (1)
- power-to-gas (1)
- prebiotic (1)
- proton therapy (1)
- protons (1)
- pullulan (1)
- recombinant expression (1)
- relative dosimetry (1)
- retention time (1)
- rubber (1)
- superabsorbent polymers (1)
- supramolecular structures (1)
- swelling properties (1)
- theory and modeling (1)
- tobacco mosaic virus (TMV) (1)
- transporters (1)
- urease (1)
- water economy (1)
- yield (1)
- α-aminoacylase (1)
- ε-lysine acylase (1)
The composition of plant biomass varies depending on the feedstock and pre-treatment conditions and influences its processing in biorefineries. In order to ensure optimal process conditions, the quantitative proportion of the main polymeric components of the pre-treated biomass has to be determined. Current standard procedures for biomass compositional analysis are complex, the measurements are afflicted with errors and therefore often not comparable. Hence, new powerful analytical methods are urgently required to characterize biomass. In this contribution, Differential Scanning Calorimetry (DSC) was applied in combination with multivariate data analysis (MVA) to detect the cellulose content of the plant biomass pretreated by Liquid Hot Water (LHW) and Organosolv processes under various conditions. Unlike conventional techniques, the developed analytic method enables the accurate quantification of monosaccharide content of the plant biomass without any previous sample preparation. It is easy to handle and avoids errors in sample preparation.
Aspergillus oryzae is an industrially relevant organism for the secretory production of heterologous enzymes, especially amylases. The activities of potential heterologous amylases, however, cannot be quantified directly from the supernatant due to the high background activity of native α-amylase. This activity is caused by the gene products of amyA, amyB, and amyC. In this study, an in vitro CRISPR/Cas9 system was established in A. oryzae to delete these genes simultaneously. First, pyrG of A. oryzae NSAR1 was mutated by exploiting NHEJ to generate a counter-selection marker. Next, all amylase genes were deleted simultaneously by co-transforming a repair template carrying pyrG of Aspergillus nidulans and flanking sequences of amylase gene loci. The rate of obtained triple knock-outs was 47%. We showed that triple knockouts do not retain any amylase activity in the supernatant. The established in vitro CRISPR/Cas9 system was used to achieve sequence-specific knock-in of target genes. The system was intended to incorporate a single copy of the gene of interest into the desired host for the development of screening methods. Therefore, an integration cassette for the heterologous Fpi amylase was designed to specifically target the amyB locus. The site-specific integration rate of the plasmid was 78%, with exceptional additional integrations. Integration frequency was assessed via qPCR and directly correlated with heterologous amylase activity. Hence, we could compare the efficiency between two different signal peptides. In summary, we present a strategy to exploit CRISPR/Cas9 for gene mutation, multiplex knock-out, and the targeted knock-in of an expression cassette in A. oryzae. Our system provides straightforward strain engineering and paves the way for development of fungal screening systems.